Outcrossing Species Research Articles

Genome-wide association mapping of latex yield and girth in Amazonian accessions of Hevea brasiliensis grown in a suboptimal climate zone

Latex yield and growth are the key complex traits in commercial rubber production. The present study is the first to report genome-wide association mapping of latex yield and girth, for 170 Amazonian accessions grown in a suboptimal area characterized by limited rainfall and a lengthy dry season. Targeted sequence enrichment to capture gene transcripts generated 14,155 high quality filtered single nucleotide polymorphisms (SNPs) of which 94.3% resided in coding regions. The rapid decay of linkage disequilibrium over physical and genetic distance found in the accessions was comparable to those previously reported for several outcrossing species. A mixed linear model detected three significant SNPs in three candidate genes involved in plant adaptation to drought stress, individually explaining 12.7–15.7% of the phenotypic variance. The SNPs identified in the study will help to extend understanding, and to support genetic improvement of rubber trees grown in drought-affected regions.

Discrimination of candidate subgenome-specific loci by linkage map construction with an S1 population of octoploid strawberry (Fragaria\u2009\xd7\u2009ananassa)

BackgroundThe strawberry, Fragaria × ananassa, is an allo-octoploid (2n = 8x = 56) and outcrossing species. Although it is the most widely consumed berry crop in the world, its complex genome structure has hindered its genetic and genomic analysis, and thus discrimination of subgenome-specific loci among the homoeologous chromosomes is needed. In the present study, we identified candidate subgenome-specific single nucleotide polymorphism (SNP) and simple sequence repeat (SSR) loci, and constructed a linkage map using an S1 mapping population of the cultivar ‘Reikou’ with an IStraw90 Axiom® SNP array and previously published SSR markers.ResultsThe ‘Reikou’ linkage map consisted of 11,574 loci (11,002 SNPs and 572 SSR loci) spanning 2816.5 cM of 31 linkage groups. The 11,574 loci were located on 4738 unique positions (bin) on the linkage map. Of the mapped loci, 8999 (8588 SNPs and 411 SSR loci) showed a 1:2:1 segregation ratio of AA:AB:BB allele, which suggested the possibility of deriving loci from candidate subgenome-specific sequences. In addition, 2575 loci (2414 SNPs and 161 SSR loci) showed a 3:1 segregation of AB:BB allele, indicating they were derived from homoeologous genomic sequences. Comparative analysis of the homoeologous linkage groups revealed differences in genome structure among the subgenomes.ConclusionsOur results suggest that candidate subgenome-specific loci are randomly located across the genomes, and that there are small- to large-scale structural variations among the subgenomes. The mapped SNPs and SSR loci on the linkage map are expected to be seed points for the construction of pseudomolecules in the octoploid strawberry.

Genetic Diversity and Population Structure of Varronia curassavica: A Medicinal Polyploid Species in a Threatened Ecosystem.

Varronia curassavica is an important medicinal species associated with the restinga, one of the most threatened coastal ecosystems of the Atlantic Forest. These circumstances call for studies aimed at estimating effective population size and gene flow to improve conservation efforts. Hence, the present study aimed to characterize the genetic diversity, ploidy level, and population structure of this species in different areas of restinga using microsatellites. Varronia curassavica was characterized as an autotetraploid, with high genetic variability, low divergence, and no significant fixation indices, indicating the absence of, or reduced, inbreeding and genetic drift in the study area. About 44% of the alleles occurred at low frequency in adults of all populations and 41% in the progenies evaluated. Gene flow was high, consistent with outcrossing species with high dispersal capacity (Nm = 4.87). The results showed no tendency toward isolation by distance. The estimated effective size indicates that the populations studied have the potential to ensure conservation of the species in the long term. The genetic variability and population structure of V. curassavica, as determined in this study, could form the foundation for activities directed toward the sustainable use of this resource and its conservation. Even though the restinga ecosystem has suffered dramatic reductions in area, this study provides evidence that this species is resilient to anthropogenic threats to its genetic integrity, since it is a polyploid with self-incompatibility mechanisms that contribute to maintaining high genetic diversity in an panmictic meta-population along the coast of Santa Catarina.

Reproductive Strategy of the Polyploid Species Varronia curassavica Jacq. in Restinga Environment.

This study aimed to elucidate the breeding strategies of Varronia curassavica, an important medicinal species associated with Brazilian restinga. This was accomplished by combining phenological and genetic data. Every 2 weeks over a period of 2 years, we measured flowering and fruiting phenology to evaluate the activity and intensity of phenophases (n = 60). We evaluated the mating system, pollen ovule ratio and genotypes from progeny and mother plants using 8 nuclear microsatellite loci. We observed flowering and fruiting of V. curassavica at low intensity throughout the entire year, but with 2 distinct peaks, one of which was seasonal, corresponding to the period of gradual increase of temperature and photoperiod. Overlapping of flowering and fruiting strategies favors gene flow among different groups of individuals and between populations by attraction of fauna throughout the year. Analysis of the mating system indicates that V. curassavica is a typical outcrossed species (t^ = 0.98; pollen/ovule ratio = 7087.50). Combining phenology with genetic studies improved our understanding of the reproductive strategies of this species. The typical outcrossing system of V. curassavica reflects the existence of functional self-incompatibility mechanisms still unaffected by changes in genetic balance by polyploidy.

Floral traits and mating system of Hibiscus trionum (Malvaceae)

Variations in floral traits and floral structures influence plant mating systems. Hibiscus trionum produces large, showy flowers typical of an outcrossing species, yet flowers are autonomously self-pollinated. In this study, we measured floral morphology, breeding system and outcrossing rate estimated by ISSR markers. Results indicate that two types of flowers were observed in H. trionum, and the type I with bigger petals appears to be much more visible to pollinators, demonstrated by than type II flowers with smaller petals. The flowers with hand pollination were closed 1h earlier than intact flowers, whether they were type I or II. The relationship between the amount of pollen deposited on the stigma and the number of seeds per capsule was highly significant, and 80 or more pollens per flower can make the mean number of seeds (mean=37) in H. trionum. Delayed selfing in H. trionum did not provide a large contribution to seed production, since reproductive assurance were only 0.025. However, successful reproduction of 72.5% flowers in the absence of pollinators suggested that selfing provides reproductive assurance during seasons, in which pollinators were limiting. The multilocus outcrossing rates in different populations varied from 0.982 to 1.200, with a mean of 1.116. Our data provide an empirical demonstration of a predominantly outcrossing species with potential delayed selfing when pollinators are absent or scarce.

Pollen-mediated gene flow from transgenic perennial creeping bentgrass and hybridization at the landscape level.

The planting of 162 ha of transgenic glyphosate-resistant creeping bentgrass (Agrostis stolonifera) near Madras, OR, USA, allowed a unique opportunity to study gene flow over time from a perennial outcrossing species at the landscape level. While conducting a four year in situ survey, we collected panicles and leaf tissue samples from creeping bentgrass and its sexually compatible species. Seeds from the panicles were planted, and seedlings were tested in the greenhouse for expression of the transgene. Gene flow via pollen was found in all four years, at frequencies of 0.004 to 2.805%. Chloroplast markers, in combination with internal transcribed spacer nuclear sequence analysis, were used to aid in identification of transgenic interspecific and intergeneric hybrid seedlings found during the testing and of established plants that could not be positively identified in the field. Interspecific transgenic hybrids produced on redtop (Agrostis gigantea) plants in situ were identified three of the four years and one intergeneric transgenic creeping bentgrass x rabbitfoot grass (Polypogon monspeliensis) hybrid was identified in 2005. In addition, we confirmed a non-transgenic creeping bentgrass x redtop hybrid in situ, demonstrating that interspecific hybrids have established in the environment outside production fields. Results of this study should be considered for deregulation of transgenic events, studies of population dynamics, and prediction of gene flow in the environment.

Unravelling wild carrot differentiation in Europe – preliminary data on a candidate gene approach

Carrot is an outcrossing species and levels of gene flow between populations, and even between wild and domesticated relatives, are expected to be high. Cases of natural hybridization and introgression of crops and wild relatives have been reported. Have these events diluted any putative habitat-adapted genotypes? In other words, can we still find a correlation between wild carrot genotypes and regional/local environment? We have chosen to start addressing this question using a member of the alternative oxidase (AOX) gene family. AOX genes seem to be linked to all kinds of abiotic and biotic stress reactions. Wild carrots were sampled in an environmental gradient across Western Europe. This gradient included sampling points with more deviating conditions, such as Sierra de Guadarrama or the central Pyrenees and the French Massif Central. Phylogenetic reconstruction on this molecular marker is to be combined with geographic, climatic, and ecological evidence. So far, the preliminary results suggest the existence of a biogeographical barrier at the Pyrenees, and higher gene diversity than initially expected. From an applied point of view, diversity of functional traits is much more relevant than species diversity. Gene transfer from wild to cultivated plants has contributed to the evolution of crop species. Providing that deterioration of genetic resources and biodiversity loss have not been drastic, gene transfer from wild plants has the potential to further contribute to a (targeted) improvement of cultivars.

Use of modern tomato breeding germplasm for deciphering the genetic control of agronomical traits by Genome Wide Association study.

A panel of 300 tomato accessions including breeding materials was built and characterized with >11,000 SNP. A population structure in six subgroups was identified. Strong heterogeneity in linkage disequilibrium and recombination landscape among groups and chromosomes was shown. GWAS identified several associations for fruit weight, earliness and plant growth. Genome-wide association studies (GWAS) have become a method of choice in quantitative trait dissection. First limited to highly polymorphic and outcrossing species, it is now applied in horticultural crops, notably in tomato. Until now GWAS in tomato has been performed on panels of heirloom and wild accessions. Using modern breeding materials would be of direct interest for breeding purpose. To implement GWAS on a large panel of 300 tomato accessions including 168 breeding lines, this study assessed the genetic diversity and linkage disequilibrium decay and revealed the population structure and performed GWA experiment. Genetic diversity and population structure analyses were based on molecular markers (>11,000 SNP) covering the whole genome. Six genetic subgroups were revealed and associated to traits of agronomical interest, such as fruit weight and disease resistance. Estimates of linkage disequilibrium highlighted the heterogeneity of its decay among genetic subgroups. Haplotype definition allowed a fine characterization of the groups and their recombination landscape revealing the patterns of admixture along the genome. Selection footprints showed results in congruence with introgressions. Taken together, all these elements refined our knowledge of the genetic material included in this panel and allowed the identification of several associations for fruit weight, plant growth and earliness, deciphering the genetic architecture of these complex traits and identifying several new loci useful for tomato breeding.

Frequency and maintenance of unreduced gametes in natural plant populations: associations with reproductive mode, life history and genome size.

Fertilization involving unreduced (2n) gametes is considered the dominant mechanism of polyploid formation in angiosperms; however, our knowledge of the prevalence of and evolutionary mechanisms maintaining 2n gametes in natural populations is limited. We hypothesize that 2n gametes are deleterious consequences of meiotic errors maintained by mutation-selection balance and should increase in species with relaxed opportunities for selection on sexual processes (asexuality), reduced efficacy of selection (asexuality, selfing) and increased genome instability (high chromosome number). We used flow cytometry to estimate male 2n gamete production in 60 populations from 24 species of Brassicaceae. We quantified variation in 2n gamete production within and among species, and examined associations with life history, reproductive mode, genome size and chromosomal number while accounting for phylogeny. Most individuals produced <2% 2n male gametes, whereas a small number had >5% (up to 85%) production. Variation in 2n gamete production was significant among species and related to reproductive system; asexual species produced significantly more 2n gametes than mixed-mating and outcrossing species. Our results, unique in their multi-species perspective, are consistent with 2n gametes being deleterious but maintained when opportunities for selection are limited. Rare individuals with elevated 2n gamete production may be key contributors to polyploid formation.

Measuring the Inefficiency of Allowing Self‐Pollinated Alfalfa Progeny into Breeding Nurseries

Although alfalfa (Medicago sativa L.) is considered an outcrossing species, the self‐pollination incidence during insect pollination can be substantial, with an observed 30% average selfing rate. Despite this, alfalfa breeders utilizing space‐plant evaluation nurseries make no attempt to discriminate self‐pollination‐derived progeny (selfs) from outcross progeny. Due to alfalfa's substantial inbreeding depression, the selfed individuals have much lower breeder utility. This study examines an alfalfa space‐plant breeding nursery in which selfed and outcross progeny had been identified using DNA markers for the following phenotypic traits: plant vigor, fall height, fall dormancy, first and second winter survival, plant lodging, and profusion of flowering. Trait means, phenotypic and additive variances, and narrow‐sense heritabilities on a halfsib mean basis were determined for selfed and outcross progeny. For most traits, inbreeding depression was severe, with approximately 80% reductions in performance. Selfed progeny for most traits had reduced phenotypic variances compared with outcross progeny. No differences in estimated additive variance and heritabilities were observed between the two types of progeny. Assuming a 30% selfing rate and a selection intensity less than or equal to 10%, we would expect to select few if any selfed progeny from a space‐plant nursery that had plant vigor phenotypic distributions observed in this study. The estimated maximum loss in genetic gain due to the presence of selfed progeny in breeding nurseries for selection intensities from 1 to 10% was estimated at a 10% genetic gain reduction. We conclude that selfed progeny in breeding nurseries are a hindrance, and if possible, they should be minimized or eliminated.

Selfing, Local Mate Competition, and Reinforcement.

Reinforcement can contribute to speciation by increasing the strength of prezygotic isolating mechanisms. Theoretical analyses over the past two decades have demonstrated that conditions for reinforcement are not unduly restrictive, and empirical investigations have documented over a dozen likely cases, indicating that it may be a reasonably common phenomenon in nature. Largely uncharacterized, however, is the diversity of biological scenarios that can create the reduced hybrid fitness that drives reinforcement. Here I examine one such scenario-the evolution of the "selfing syndrome" (a suite of characters including reductions in flower size and in nectar, pollen, and scent production) in highly selfing plant species. Using a four-locus model, where the loci are (1) a discrimination locus, (2) a target-of-discimination locus, (3) a pollen-production locus, and (4) a selfing-rate locus, I determine the conditions under which this syndrome can favor reinforcement, an increase in discrimination through change at locus 1, in an outcrossing species that experiences gene flow from a highly selfing species. In the absence of both linkage disequilibrium between loci and pollen discounting, reinforcement can occur, but only in a very small fraction of the parameter combinations examined. Moderate linkage ([Formula: see text]) between one pair of loci increases this fraction moderately, depending on which two loci are linked. Pollen discounting (a reduction in pollen exported to other plants due to increased selfing), by contrast, can increase the fraction of parameter combinations that result in reinforcement substantially. The evolution of reduced pollen production in highly selfing species thus facilitates reinforcement, especially if substantial pollen discounting is associated with selfing.

Maintenance of mixed mating after the loss of self-incompatibility in a long-lived perennial herb.

Many hermaphroditic plants avoid self-fertilization by means of a molecular self-incompatibility (SI) system, a complex trait that is difficult to evolve but relatively easy to lose. Loss of SI is a prerequisite for an evolutionary transition from obligate outcrossing to self-fertilization, which may bring about rapid changes in the genetic diversity and structure of populations. Loss of SI is also often followed by the evolution of a 'selfing syndrome', with plants having small flowers, little nectar and few pollen grains per ovule. Here, we document the loss of SI in the long-lived Spanish toadflax Linaria cavanillesii, which has led to mixed mating rather than a transition to a high rate of selfing and in which an outcrossing syndrome has been maintained. We performed crosses within and among six populations of L. cavanillesii in the glasshouse, measured floral traits in a common-garden experiment, performed a pollen-limitation experiment in the field and conducted population genetic analyses using microsatellites markers. Controlled crosses revealed variation in SI from fully SI through intermediate SI to fully self-compatible (SC). Flowers of SC individuals showed no evidence of a selfing syndrome. Although the SC population of L. cavanillesii had lower within-population genetic diversity than SI populations, as expected, population differentiation among all populations was extreme and represents an FST outlier in the distribution for both selfing and outcrossing species of flowering plants. Together, our results suggest that the transition to SC in L. cavanillesii has probably been very recent, and may have been aided by selection during or following a colonization bottleneck rather than in the absence of pollinators. We find little indication that the transition to SC has been driven by selection for reproductive assurance under conditions currently prevailing in natural populations.

A high-density, multi-parental SNP genetic map on apple validates a new mapping approach for outcrossing species.

Quantitative trait loci (QTL) mapping approaches rely on the correct ordering of molecular markers along the chromosomes, which can be obtained from genetic linkage maps or a reference genome sequence. For apple (Malus domestica Borkh), the genome sequence v1 and v2 could not meet this need; therefore, a novel approach was devised to develop a dense genetic linkage map, providing the most reliable marker-loci order for the highest possible number of markers. The approach was based on four strategies: (i) the use of multiple full-sib families, (ii) the reduction of missing information through the use of HaploBlocks and alternative calling procedures for single-nucleotide polymorphism (SNP) markers, (iii) the construction of a single backcross-type data set including all families, and (iv) a two-step map generation procedure based on the sequential inclusion of markers. The map comprises 15 417 SNP markers, clustered in 3 K HaploBlock markers spanning 1 267 cM, with an average distance between adjacent markers of 0.37 cM and a maximum distance of 3.29 cM. Moreover, chromosome 5 was oriented according to its homoeologous chromosome 10. This map was useful to improve the apple genome sequence, design the Axiom Apple 480 K SNP array and perform multifamily-based QTL studies. Its collinearity with the genome sequences v1 and v3 are reported. To our knowledge, this is the shortest published SNP map in apple, while including the largest number of markers, families and individuals. This result validates our methodology, proving its value for the construction of integrated linkage maps for any outbreeding species.

High Correlated Paternity Leads to Negative Effects on Progeny Performance in Two Mediterranean Shrub Species.

Anthropogenic habitat deterioration can promote changes in plant mating systems that subsequently may affect progeny performance, thereby conditioning plant recruitment for the next generation. However, very few studies yet tested mating system parameters other than outcrossing rates; and the direct effects of the genetic diversity of the pollen received by maternal plants (i.e. correlated paternity) has often been overlooked. In this study, we investigated the relation between correlated paternity and progeny performance in two common Mediterranean shrubs, Myrtus communis and Pistacia lentiscus. To do so, we collected open-pollinated progeny from selected maternal plants, calculated mating system parameters using microsatellite genotyping and conducted sowing experiments under greenhouse and field conditions. Our results showed that some progeny fitness components were negatively affected by the high correlated paternity of maternal plants. In Myrtus communis, high correlated paternity had a negative effect on the proportion and timing of seedling emergence in the natural field conditions and in the greenhouse sowing experiment, respectively. In Pistacia lentiscus, seedling emergence time under field conditions was also negatively influenced by high correlated paternity and a progeny survival analysis in the field experiment showed greater mortality of seedlings from maternal plants with high correlated paternity. Overall, we found effects of correlated paternity on the progeny performance of Myrtus communis, a self-compatible species. Further, we also detected effects of correlated paternity on the progeny emergence time and survival in Pistacia lentiscus, an obligate outcrossed species. This study represents one of the few existing empirical examples which highlight the influence that correlated paternity may exert on progeny performance in multiple stages during early seedling growth.

Conserved imprinting associated with unique epigenetic signatures in the Arabidopsis genus.

In plants, imprinted gene expression occurs in endosperm seed tissue and is sometimes associated with differential DNA methylation between maternal and paternal alleles(1). Imprinting is theorized to have been selected for because of conflict between parental genomes in offspring(2), but most studies of imprinting have been conducted in Arabidopsis thaliana, an inbred primarily self-fertilizing species that should have limited parental conflict. We examined embryo and endosperm allele-specific expression and DNA methylation genome-wide in the wild outcrossing species Arabidopsis lyrata. Here we show that the majority of A. lyrata imprinted genes also exhibit parentally biased expression in A. thaliana, suggesting that there is evolutionary conservation in gene imprinting. Surprisingly, we discovered substantial interspecies differences in methylation features associated with paternally expressed imprinted genes (PEGs). Unlike in A. thaliana, the maternal allele of many A. lyrata PEGs was hypermethylated in the CHG context. Increased maternal allele CHG methylation was associated with increased expression bias in favour of the paternal allele. We propose that CHG methylation maintains or reinforces repression of maternal alleles of PEGs. These data suggest that the genes subject to imprinting are largely conserved, but there is flexibility in the epigenetic mechanisms employed between closely related species to maintain monoallelic expression. This supports the idea that imprinting of specific genes is a functional phenomenon, and not simply a byproduct of seed epigenomic reprogramming.

Functional polymorphism in lycopene beta-cyclase gene as a molecular marker to predict bixin production in Bixa orellana L. (achiote)

Bixin is an apocarotenoid obtained from the seed aril of Bixa orellana L., a tropical plant known as achiote in Mexico. This compound is the second most commonly used natural colouring for food and pharmaceutical industries. B. orellana is an outcrossing species that displays high genetic variability. Recently, the colour traits of sexual organs were associated with the biosynthesis and accumulation of bixin in mature seeds. Herein, we describe a new approach for genotype–phenotype association by surveying lycopene beta-cyclase (Boβ-LCY1) gene variation in sixteen achiote accessions divided into three groups according to contrasting traits, such as flower colour, fruit colour and bixin production. Using a combination of single-strand conformational polymorphism techniques and the sequencing of polymorphic bands, we identified several single-nucleotide polymorphisms that divided the accessions into three haplotypes. Surprisingly, we observed that these three haplotypes were consistent with the same three groups previously characterized by phenotypic traits. We derived a putative sequence for the Boβ-LCY1 gene and surveyed the variations in this sequence. The heterozygosity of Boβ-LCY1 alleles resulted in a higher bixin content, likely associated with heterosis for this metabolite. These findings augment the toolbox available for the selection and genetic improvement of B. orellana and provide a reliable phenotype–genotype association method for commercial varietal selection, contributing to the development of laboratory techniques to identify desirable traits of commercial plant species.

QTL mapping including codominant SNP markers with ploidy level information in a sugarcane progeny

AbstractQuantitative trait locus (QTL) mapping contributes to sugarcane (Saccharum spp.) breeding programs by providing information about the genetic effects, positioning and number of QTLs. Combined with marker-assisted selection, it can help breeders reduce the time required to develop new sugarcane varieties. We performed a QTL mapping study for important agronomic traits in sugarcane using the composite interval mapping method for outcrossed species. A new approach allowing the 1:2:1 segregation ratio and different ploidy levels for SNP markers was used to construct an integrated genetic linkage map that also includes AFLP and SSR markers. Were used 688 molecular markers with 1:1, 3:1 and 1:2:1 segregation ratios. A total of 187 individuals from a bi-parental cross (IACSP95-3018 and IACSP93-3046) were assayed across multiple harvests from two locations. The evaluated yield components included stalk diameter (SD), stalk weight (SW), stalk height (SH), fiber percentage (Fiber), sucrose content (Pol) and soluble solid content (Brix). The genetic linkage map covered 4512.6 cM and had 118 linkage groups corresponding to 16 putative homology groups. A total of 25 QTL were detected for SD (six QTL), SW (five QTL), SH (four QTL), Fiber (five QTL), Pol (two QTL) and Brix (three QTL). The percentage of phenotypic variation explained by each QTL ranged from 0.069 to 3.87 %, with a low individual effect because of the high ploidy level. The mapping model provided estimates of the segregation ratio of each mapped QTL (1:2:1, 3:1 or 1:1). Our results provide information about the genetic organization of the sugarcane genome and constitute the first step toward a better dissection of complex traits.

Reinventing Potato as a Diploid Inbred Line–Based Crop

The third most important food crop worldwide, potato (Solanum tuberosum L.) is a tetraploid outcrossing species propagated from tubers. Breeders have long been challenged by polyploidy, heterozygosity, and asexual reproduction. It has been assumed that tetraploidy is essential for high yield, that the creation of inbred potato is not feasible, and that propagation by seed tubers is ideal. In this paper, we question those assumptions and propose to convert potato into a diploid inbred line–based crop propagated by true seed. Although a conversion of this magnitude is unprecedented, the possible genetic gains from a breeding system based on inbred lines and the seed production benefits from a sexual propagation system are too large to ignore. We call on leaders of public and private organizations to come together to explore the feasibility of this radical and exciting new strategy in potato breeding.

Survey of genetic diversity and seed germination rates of the southeastern endemic Symphyotrichum georgianum (Alexander) G.L. Nesom (Asteraceae) from large and small populations1,2

Abstract Single nucleotide polymorphic (SNP) markers were used to characterize the genetic structure of Symphyotrichum georgianum (Alexander) G.L. Nesom, known more commonly as the Georgia aster, from 26 populations across the species' range. This species is considered vulnerable (G3) and was a candidate for listing under the Endangered Species Act. We sampled large (> 500 stems) and small (< 100 stems) populations from across the entire range for the species. Next generation sequencing was used to assess 36 SNP markers for pooled population samples, consisting of equal amounts of genomic DNA from 30 individual stems sampled across each population. Most of the genetic variation was partitioned within and less among populations, which is consistent with perennial outcrossing species. There was a significant (r = 0.506, P < 0.001) positive association between geographic distance and genetic distance among populations, indicating increasing genetic distance (genetic dissimilarity) with increasing geographic ...

Kunzea pomifera(muntries): selection validation and evaluation of important horticultural traits

The native food industry in Australia has been developed for nearly 40 years. This industry is focussed on 13 crops, including quandong and muntries (or muntrie berries). There are approximately 20 selections of muntries such as 'Rivoli Bay' and 'Maarten's Favourite'. 'Rivoli Bay', the industry standard selection, was chosen for evaluation at Mount Pleasant, South Australia, Australia from February to March 2014. Assessment was performed on five trellised 'Rivoli Bay' plants. Methods for the evaluation of muntries fruit size and fruit quality characteristics (texture, total soluble solids and total phenolic content) were developed from existing methods for other fruits. Variability between the five 'Rivoli Bay' plants was higher than expected for all assessed fruit characteristics, given that plants are clonally propagated, which is possibly due to an environmental effect. Results obtained for n=3 technical replicates produced low standard deviation suggesting the methods developed for assessing muntries fruit characteristics can be used in the future. To determine genetic diversity and identify cultivars, molecular markers are commonly used. We applied this approach to muntries and primers for moleculars were designed in introns or at intron-exon boundaries of eight Myb gene fragments that include one or two short introns. Three PCR fragments sequenced contained polymorphic sites between the selections. These polymorphisms included insertion/deletions (Indels) and a clear single-nucleotide polymorphism (SNP) where several individuals were heterozygous for the SNP. Our data suggested that heterozygosity is common in 'Rivoli Bay' as expected for a preferentially outcrossing species.